Direct Cellular Effects of Blood Coagulation Proteases

《独立之路》(K99/Roo)职业发展奖申请的主要科学目标 是通过研究新的研究来了解血栓和炎症性疾病的分子病理生理学 维生素K依赖的凝血酶的细胞保护作用机制。此应用程序 最初侧重于膜受体在调节细胞蛋白C途径中的作用 随后对凝血酶的细胞保护活性的新机制进行了探索。少校 应聘者的职业发展目标是扩大其所需的技术和学术经验 成功转型为独立调查员。这些研究将提供机会和坚实的 为成功申请未来NIH R01分子机制方面的独立资金奠定基础 研究集中在凝血和炎症的十字路口。关于泛函的新假设 凝血酶的细胞保护作用的蛋白质组学将用生化和 细胞生物学方法。拟议研究的临床和治疗意义从 在大型临床试验中，激活的蛋白C(ARC)而不是其他抗凝剂降低了重症患者的死亡率 并暗示APC的抗凝活性和直接活性的独特组合 CELES是APC成功的基础。我已发表的作品和未发表的初步数据直接导致了 并为我的假设提供了强有力的支持。在检验这些假设时，我提出：1) 为了表征血栓调节蛋白之间内皮细胞膜受体复合体的形成， APC生成和APC所需的内皮蛋白C受体和蛋白酶激活受体-1 对细胞的直接影响；2)阐明血小板的潜在有益和有害功能特性 APC生成的因子4和APC对细胞的直接影响；3)确定新的主题和机制 通过探讨APC和fV11a对细胞保护作用的异同 APC和fV11a抗细胞凋亡活性；4)确定美佐托品是否具有抗细胞凋亡作用 如果这一活动需要辅因子依赖和PAR依赖的机制。如果 建议的研究是成功的，它们将增加我们的知识，并可能导致改善治疗 血栓形成、细胞凋亡和炎症导致的各种疾病。

The major scientific goal of this Pathway to Independence (K99/ROO) Career Development Award application is to understand the molecular pathophysiology of thrombotic and inflammatory disorders by studying novel mechanisms for cytoprotective actions of vitamin K-dependent coagulation proteases. This application focuses initially on the role of membrane receptors in the regulation of the cellular protein C pathway and later on the exploration of novel mechanisms for cytoprotective activities of coagulation proteases. The major career development goal of the applicant is to expand his technical and academic experience required for a successful transition into an independent investigator. These studies will provide the opportunity and solid basis to apply successfully for future independent NIH R01 funding focused on the molecular mechanistic studies centered on the crossroads of coagulation and inflammation. Novel hypotheses on the functional proteomics of cytoprotective actions by blood coagulation proteases will be tested using biochemical and cellular biology methods. The clinical and therapeutic implications of the proposed studies are clear from the large clinical trials, where activated protein C (ARC), but not other anticoagulants reduced mortality in severe sepsis patients and implied that the unique combination of APC's anticoagulant activity and direct activity on cells is the basis for APC's success. My published work and unpublished preliminary data lead directly to the proposed studies and provide strong support for my hypotheses. In testing these hypotheses, I propose: 1) To characterize the formation of endothelial cell membrane receptor complexes between thrombomodulin, endothelial protein C receptor and protease activated receptor-1 required for APC generation and APC's direct effects on cells; 2) To clarify the potential beneficial and detrimental functional properties of platelet factor 4 for APC generation and APC's direct effects on cells; 3) To identify novel themes and mechanisms for APC and fVlla cytoprotective actions on cells by exploration of the similarities and differences between APC and fVlla anti-apoptotic activities; and 4) To establish whether meizothrombin has anti-apoptotic activity, as predicted, and if this activity requires cofactor-dependent and PAR-dependent mechanisms. If the proposed studies are successful, they will increase our knowledge and may lead to improved treatment of a variety of disorders in which thrombosis, apoptosis and inflammation contribute to pathogenesis.